Use of markers of undifferentiated pluripotent stem cell

• US 8,906,678 B2
• Filed: 03/10/2011
• Issued: 12/09/2014
• Est. Priority Date: 03/10/2010
• Status: Active Grant

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Agency For Science Technology and Research
• Inventors: Choo, Boon Hwa Andre, Fong, Wey Jia
• Primary Examiner(s): Singh, Anoop
• Assistant Examiner(s): Montanari, David A
• Application Number:
  US13/583,546
• Publication Number:
  US 20130115623A1
• Time in Patent Office: 1,370 Days
• Field of Search: 435/325, 435/347, 435/366, 435/354
• US Class Current: 435/325
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07D 409/14 containing three or more hetero rings
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The disclosure relates to methods of binding and identifying undifferentiated pluripotent stem cells and particularly, although not exclusively, to use of binding moieties which bind to PHB on the surface of undifferentiated pluripotent stem cells, such as PHB-binding peptides, and to methods for depleting undifferentiated stem cells from a sample.

Claims

1. A method of isolating a human or mouse embryonic stem (ES) cells or induced pluripotent stem (iPS) cells in a heterogeneous sample containing human or mouse ES cells or iPS cells and cells that have undergone or are undergoing differentiation from human or mouse pluripotent ES cells or iPS cells, comprising:

(a) obtaining a heterogeneous sample containing human or mouse ES cells or iPS cells and cells that have undergone or are undergoing differentiation from human or mouse pluripotent ES cells or iPS cells,

(b) contacting the heterogeneous sample of step (a) with an antibody which binds prohibitin (PHB),

(c) identifying ES iPS cells in the sample of step (b) which are bound to the PHB antibody; and

(d) isolating the ES or iPS cells from step (c) that express PHB on their cell surface.

2. A method of enriching a human or mouse embryonic stem (ES) cells or induced pluripotent stem (iPS) cells in a heterogeneous sample containing human or mouse ES cells or iPS cells and cells that have undergone or are undergoing differentiation from human or mouse pluripotent ES cells or iPS cells, comprising:

(a) obtaining a heterogeneous sample containing human or mouse ES cells or iPS cells and cells that have undergone or are undergoing differentiation from human or mouse pluripotent ES cells or iPS cells,

(b) contacting the heterogeneous sample of step (a) with an antibody which binds prohibitin (PHB),

(c) identifying ES iPS cells in the sample of step (b) which are bound to the PHB antibody; and

(d) enriching the ES or iPS cells from step (c) that express PHB on their cell surface.

3. A method of destroying human or mouse ES or iPS cells in a heterogeneous sample containing human or mouse ES or iPS cells and cells that have undergone or are undergoing differentiation from human or mouse ES or iPS cells, comprising:

(a) obtaining a heterogeneous sample containing human or mouse pluripotent ES or iPS and cells that have undergone or are undergoing differentiation from human or mouse ES or iPS cells,

(b) contacting the heterogeneous sample of step (a) with a PHB targeting pro-apoptotic peptide comprising the amino acid sequence set forth in SEQ ID NO;

3, and

(c) destroying the PHB-expressing human or mouse pluripotent ES or iPS cells in the sample of step (b).